Ling Zhang1, Mingdong Zhao2, Fang Jiao1, Xiaoyan Xu3, Xianbing Liu1, Yuzhu Jiang1, Haixia Zhang1, Xiaoxuan Ou1, Xuemei Hu4. 1. Department of Immunology, Binzhou Medical University, No. 346 Guan-Hai Road, Lai-shan, Yantai, Shandong 264003, PR China. 2. Department of Radiology, Binzhou Affiliated Hospital of Binzhou Medical University, Binzhou, Shandong, PR China. 3. Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, Shandong, PR China. 4. Department of Immunology, Binzhou Medical University, No. 346 Guan-Hai Road, Lai-shan, Yantai, Shandong 264003, PR China; Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, Shandong, PR China. Electronic address: xue-mei-hu@163.com.
Abstract
OBJECTIVE: To assess whether interferon gamma (IFN-γ) secreted by decidual natural killer (dNK) cells at the maternal-fetal interface is involved in the apoptosis of trophoblast cells (trophoblasts) following Toxoplasma gondii infection. METHODS: Human dNK cells were infected with T. gondii and then co-cultured with trophoblasts. The infected co-cultured cells were treated with or without IFN-γ neutralizing antibodies. Uninfected co-cultured cells were used as controls. The level of IFN-γ in the supernatant of co-culture was measured by ELISA and the apoptosis of trophoblasts was analyzed by flow cytometry. The expression of caspase 3 and caspase 8 of trophoblasts cells was determined by Western blotting and real-time RT-PCR. RESULTS: The levels of IFN-γ were increased at <24h following T. gondii infection and were maintained thereafter. Caspase 3, caspase 8, and the apoptosis of trophoblasts co-cultured with dNK cells were increased compared with the control. Meanwhile, IFN-γ, caspase 3, caspase 8, and trophoblast apoptosis were up-regulated upon increased ratios of dNK cells to trophoblasts. Compared to the infected group, the levels of IFN-γ, caspase 3, caspase 8, and trophoblast apoptosis were significantly decreased upon treatment with the IFN-γ neutralizing antibody. IFN-γ levels were correlated positively with the apoptosis of trophoblasts (r=0.7163, p < 0.01). CONCLUSIONS: The levels of IFN-γ secreted by dNK cells at the maternal-fetal interface were closely correlated with the apoptosis of trophoblasts upon T. gondii infection. The apoptosis of trophoblasts induced by the increase in IFN-γ depended on the caspase pathway, which may contribute to the abnormal pregnancy outcomes that occur with T. gondii infection.
OBJECTIVE: To assess whether interferon gamma (IFN-γ) secreted by decidual natural killer (dNK) cells at the maternal-fetal interface is involved in the apoptosis of trophoblast cells (trophoblasts) following Toxoplasma gondii infection. METHODS:HumandNK cells were infected with T. gondii and then co-cultured with trophoblasts. The infected co-cultured cells were treated with or without IFN-γ neutralizing antibodies. Uninfected co-cultured cells were used as controls. The level of IFN-γ in the supernatant of co-culture was measured by ELISA and the apoptosis of trophoblasts was analyzed by flow cytometry. The expression of caspase 3 and caspase 8 of trophoblasts cells was determined by Western blotting and real-time RT-PCR. RESULTS: The levels of IFN-γ were increased at <24h following T. gondii infection and were maintained thereafter. Caspase 3, caspase 8, and the apoptosis of trophoblasts co-cultured with dNK cells were increased compared with the control. Meanwhile, IFN-γ, caspase 3, caspase 8, and trophoblast apoptosis were up-regulated upon increased ratios of dNK cells to trophoblasts. Compared to the infected group, the levels of IFN-γ, caspase 3, caspase 8, and trophoblast apoptosis were significantly decreased upon treatment with the IFN-γ neutralizing antibody. IFN-γ levels were correlated positively with the apoptosis of trophoblasts (r=0.7163, p < 0.01). CONCLUSIONS: The levels of IFN-γ secreted by dNK cells at the maternal-fetal interface were closely correlated with the apoptosis of trophoblasts upon T. gondii infection. The apoptosis of trophoblasts induced by the increase in IFN-γ depended on the caspase pathway, which may contribute to the abnormal pregnancy outcomes that occur with T. gondii infection.